Human embryonic stem cells (hESCs) are pluripotent cells derived from the inner cell mass of pre-implantation embryos (Thomson J, et al., Science 282:1145-1147 (1998)). Similar to mouse embryonic stem cells (mESCs), they can be expanded to large numbers while maintaining their potential to differentiate into various somatic cell types of all three germ layers (Thomson et al., supra; Reubinoff B, et al., Nat. Biotech. 18:399 (2000); Thomson J & Odorico J, Trends Biotech. 18:53-57 (2000); and Amit M, et al., Dev. Biol. 227:271-278 (2000)). The in vitro differentiation of embryonic stem cells (ESCs) provides new perspectives for studying the cellular and molecular mechanisms of early development and the generation of donor cells for transplantation therapies. Indeed, mESCs have been found to differentiate in vitro to many clinically relevant cell types, including hematopoietic cells (Wiles M & Keller G, Development 111:259-267 (1991)), cardiomyocytes (Klug M, et al., J. Clin. Invest. 98:216-224 (1996)), insulin-secreting cells (Soria B, et al., Diabetes 49:157-162 (2000)), and neurons and glia (Bain G, et al., Dev. Biol. 168:342-357 (1995); Okabe S, et al., Mech. Dev. 59:89-102 (1996); Mujtaba T, et al., Dev. Biol. 214:113-127 (1999); and Brustle O, et al., Science 285:754-756 (1999)). Following transplantation into the rodent central nervous system (CNS), ESC-derived neural precursors have been shown to integrate into the host tissue and, in some cases, yield functional improvement (McDonald J, et al., Nat. Med. 5:1410-1412 (1999)). A clinical application of hESCs would require the generation of highly purified donor cells for specific tissues and organs.
Needed in the art is a simple, yet efficient, strategy for the isolation of transplantable neural and motor neuron precursors from differentiating human ES cell cultures.